The Effect Of P-gp Function And Expression At Ischemia Injuryed Rbmecs

Objective: To investigate the effect of P-gp function and expression at ischemia injuryed rBMECs.Methods: Prepare the primary cultures of rBMECs、astrocyte、neuron cells and construct BBB models in vitro with transwell.The cytotoxicity toward rBMECs was induced by different concentration of Na2S2O4. We make sure what is the most approptiate concentration of Na2SiO4,to induce celluar injuries by observing cell morphology through inverted microscope and detecting the survival rate at the time of6h with MTT. Transendothelial electrical resistance (TEER) and Papp of NaF were measured to evaluate the quality of BBB models, The Papp of Rh123were measured to evaluate paracellular transport reflecting function. The levels of P-gp in brain endothelial cells cultured in the presence of astrocytes and neuron cells, were determined by Western blot and immunofluorescence with living cells high content imaging system.Results:!.At being cultured with Na2S2O4for6hours, the survival rate,TEER and the intracellular accumulation of NaF in rBMECs and Papp of NaF can not be significantly reduced, Interendothelial tight junction was integral at6h.2.Being cultured with NaaS2O4for24hours,at0-6hours the Papp of NaF received no influence,got higher after6hours.As compared to Oh the Papp of Rhl23showed lower at the first4h and got higher after6hours.3.The expression of P-gp can be improved at6h and significantly reduced after6hours.Conclusions: The results of the experiment in vitro demonsterate that the function and expression of P-gp at ischemia injuryed rBMECs rised firstly and got lower later, which provided experiment basis for the further study of drug therapy time window of the central nervous system disease.